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My research interests
Infection biology, Microbiology, cancer biology
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In Topic: Western blotting
23 January 2013 - 06:16 PM
thank you very much for your kind reply, and i will work on it..
In Topic: Western blotting
22 January 2013 - 09:08 PM
thanks to every one for your valuable reply.
actually my primary ab - 1:2000, secondary - 1:3000, and I am using 10% skim milk in 0.05% Tween 20 in PBS to dilute the abs.
actually my primary ab - 1:2000, secondary - 1:3000, and I am using 10% skim milk in 0.05% Tween 20 in PBS to dilute the abs.
In Topic: Drug recrystallize when i added to DMEM
31 August 2012 - 01:33 AM
Dear xabi,
thank you for your reply.
First of all, I made 200 mM stock piperine solution with DMSO. Then I made 2 mM piperine working stock with 3% Tween80. Then after made piperine working stock with 3 % Tween, i would like to filter sterilization instead of Autoclave. because some of my friends suggest me its better to filter the Tween instead of autoclaving.
then from filtered working stock, i would like to add the to the cells for MTT assay. the maximum final concentration of the tween is 0.1 % and maximum DMSO concentration is below 0.01 %..
this is the way i plan to do my assay.. in your view, is that any thing to change in plan ?
Looking for your kind reply.
micronagu
thank you for your reply.
First of all, I made 200 mM stock piperine solution with DMSO. Then I made 2 mM piperine working stock with 3% Tween80. Then after made piperine working stock with 3 % Tween, i would like to filter sterilization instead of Autoclave. because some of my friends suggest me its better to filter the Tween instead of autoclaving.
then from filtered working stock, i would like to add the to the cells for MTT assay. the maximum final concentration of the tween is 0.1 % and maximum DMSO concentration is below 0.01 %..
this is the way i plan to do my assay.. in your view, is that any thing to change in plan ?
Looking for your kind reply.
micronagu
In Topic: Drug recrystallize when i added to DMEM
30 August 2012 - 08:02 PM
Dear Xabi,
thank you very much for your valuable suggestion, i tried that, my drug was dissolved in 3 % Tween 80.
now i have several doubts to ask,
1. Can i autoclave the tween 80?
2. if is it possible to autoclave, may i prepare 3 % tween with DMEM for dissolve the drug ?
3. If it is not possible to auticlave tween 80, can i filter sterlize the 3 % tween 80 ?
looking for your kind reply.
thank you,
micronagu
thank you very much for your valuable suggestion, i tried that, my drug was dissolved in 3 % Tween 80.
now i have several doubts to ask,
1. Can i autoclave the tween 80?
2. if is it possible to autoclave, may i prepare 3 % tween with DMEM for dissolve the drug ?
3. If it is not possible to auticlave tween 80, can i filter sterlize the 3 % tween 80 ?
looking for your kind reply.
thank you,
micronagu
In Topic: HEK293T cells are too fragile??
29 August 2012 - 10:21 PM
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yes, HEK 293 cell line is weakly attached to the surface, and i am using culture dish instead of flask. while changing the medium, aspirate the medium very slow rate, for example i am aspirating the medium by using 200 microL yellow tips, and adding medium as slow rate as possible.
during the trypsinization, i just gave only 30 seconds for detachment...
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